Anatomic variations in branching patterns of the axillary artery: a multidetector-row computed tomography angiography study.
Anatomic variations in branching pattern of axillary artery (AxA) are common and typically involve subscapular artery (SsA) and posterior circumflex humeral artery (PCHA). Several skin and muscle flaps are based on the branches of AxA. Furthermore, these branches are frequently used as recipient vessels in functioning free muscle transfers for upper extremity reconstruction and in breast reconstruction. Accurate knowledge of the normal anatomy and variations in branching pattern of AxA is of significant clinical importance for the reconstructive microsurgeon. The purpose of this article is to report the variable branching pattern of AxA based on multidetector-row computed tomography angiography study of 62 upper extremities. The thoracoacromial artery consistently originated from the first or second part of AxA. The classic origin and branching patterns of SsA and PCHA were observed in 21 cases (33.9%). Anatomic variations of SsA and PCHA were observed in 41 upper extremities (66.1%). In addition to the classic pattern, five distinct variations were noted.


Fusion for low-grade adult isthmic spondylolisthesis: a systematic review of the literature.
The objective of this study was to evaluate which fusion technique provides the best clinical and radiological outcome for adult low-grade lumbar isthmic spondylolisthesis, and to assess the overall clinical and radiological outcome of each fusion technique. A systematic review was performed. Medline, Embase, Current Contents, and Cochrane databases as well as reference lists of selected articles were searched. Randomised controlled trials (RCTs) were used to evaluate the best treatment; controlled studies and non-controlled studies were used to determine the outcomes after surgery. Two independent reviewers evaluated the studies with the methodological checklists of van Tulder and Jadad for the randomised studies and of Cowley for the non-randomised studies. The search resulted in 684 references and eventually 29 studies met the inclusion criteria, of which eight were RCTs, four were prospective, and 17 were retrospective case series. Ten of the case series did not clearly identify consecutive patient selection. All the eight RCTs evaluated the effect of different techniques of posterolateral fusion (PLF). Evidence was found that the PLF was superior to non-operative treatment (exercise). Circumferential fusion was compared to PLF, but no difference could be found. PLF with or without instrumentation was evaluated in three studies, but no benefits from additional instrumentation were found. Other comparisons within PLF showed no effect of decompression, alternative instrumentation, or bone graft substitute. The 21 case series included 24 patient groups. PLF was used in 15 groups, good or excellent clinical outcome varied from 60 to 98% and fusion rate varied from 81 to 100%. Anterior interbody fusion was used in five groups, good or excellent clinical outcome varied from 85 to 94% and fusion rate varied from 47 to 90%. Posterior interbody fusion was used in two groups, good or excellent clinical outcome was 45% and fusion rate was 80 and 95%, respectively. Reduction, loss of reduction, and lordotic angles before and after the treatment was reported in only four studies. Average reduction achieved was 12.3%, average loss of reduction at follow-up was 5.9%. Preoperative lordotic angles were too heterogeneous to pool the results. Adjacent segment degeneration was not reported in any of the publications. A wide variety of complications were reported in 18 studies and included neurological complications, instrument failure, and infections. Fusion for low-grade isthmic spondylolisthesis has better outcomes than non-operative treatment. The current study could not identify the best surgical technique (PLF, PLIF, ALIF, instrumentation) to perform the fusion. However, instrumentation and/or decompression may play a beneficial role in the modern practice of reduction and fusion for low-grade isthmic spondylolisthesis, but there are no studies yet available to confirm this. The outcomes of fusion are generally good, but reports vary widely.


Expression Pattern of ERF Gene Family under Multiple Abiotic Stresses in Populus simonii × P. nigra.
Identification of gene expression patterns of key genes across multiple abiotic stresses is critical for mechanistic understanding of stress resistance in plant. In the present study, we identified differentially expressed genes (DEGs) in di-haploid Populus simonii × P. nigra under respective stresses of NaCl, KCl, CdCl2, and PEG. On the basis of RNA-Seq, we detected 247 DEGs that are shared by the four stresses in wild type poplar, and mRNA abundance of the DEGs were validated in transgenic poplar overexpressing ERF76 gene by RNA-Seq and RT-qPCR. Results from gene ontology analysis indicated that these genes are enriched in significant pathways, such as phenylpropanoid biosynthesis, phenylalanine metabolism, starch and sucrose metabolism, and plant hormone signal transduction. Ethylene response factor (ERF) gene family plays significant role in plant abiotic stress responses. We also investigated expression pattern of ERF gene family under the four stresses. The ERFs and DEGs share similar expression pattern across the four stresses. The transgenic poplar is superior to WT in morphologic, physiological and biochemical traits, which demonstrated the ERF76 gene plays a significant role in stress resistance. These studies will give a rise in understanding the stress response mechanisms in poplar.


Epidemiology of human infections with highly pathogenic avian influenza A(H7N9) virus in Guangdong, 2016 to 2017.
We describe the epidemiology of highly pathogenic avian influenza (HPAI) A(H7N9) based on poultry market environmental surveillance and laboratory-confirmed human cases (n = 9) in Guangdong, China. We also compare the epidemiology between human cases of high- and low-pathogenic avian influenza A(H7N9) (n = 51) in Guangdong. Case fatality and severity were similar. Touching sick or dead poultry was the most important risk factor for HPAI A(H7N9) infections and should be highlighted for the control of future influenza A(H7N9) epidemics.


A Cost-effectiveness Analysis of Brentuximab Vedotin in Relapsed or Refractory Systemic Anaplastic Large Cell Lymphoma.
Objective: To evaluate the cost-effectiveness of brentuximab vedotin in patients with R/R sALCL from a UK NHS perspective. Methods: A partitioned survival model used clinical outcomes for brentuximab vedotin from the pivotal phase-2 single-arm trial of brentuximab vedotin in 58 patients with R/R sALCL (SG035-0004; NCT00866047), over a lifetime (30-year) time horizon. Comparison with conventional chemotherapy was based on data from the Canadian British Columbia Cancer Agency registry from 40 patients starting salvage chemotherapy after front-line treatment between 1980 and 2012. Survival was extrapolated using parametric distributions, with brentuximab vedotin risk after the trial period assumed equal to conventional chemotherapy. Other modelling assumptions were based on a systematic literature review and clinical expert opinion. Results: Based on statistical extrapolation, brentuximab vedotin was associated with 3.1 years longer duration in the progression-free survival health state and an overall survival improvement of 5.4 years, prior to discounting. In addition, brentuximab vedotin was associated with 2.5 quality-adjusted life years (QALYs) gained at a total incremental cost of £88 556, resulting in an incremental cost-effectiveness ratio (ICER) of approximately £35 400. Sensitivity analyses of alternative model assumptions provided ICERs ranging from approximately £28 100 to £61 900. Comparing only first-line salvage patients reduced the ICER to £26 800 per QALY gained. Conversely, considering only patients with Eastern Corporative Oncology Group performance status of 0 or 1 increased the ICER to approximately £38 200. At a willingness-to-pay threshold of £50 000, the estimated probability that brentuximab vedotin is cost-effective compared with conventional chemotherapy was 86.5%. Conclusion: Compared to conventional chemotherapy, and considering the full survival period, brentuximab vedotin may provide a valuable treatment choice for patients with R/R sALCL, a population with limited therapeutic options.


Trend in mortality from a recent measles outbreak in Cameroon: a retrospective analysis of 223 measles cases in the Benakuma Health District.
Measles is a highly contagious viral infection with high mortality in poorly vaccinated regions. We sought to establish the trend in mortality and the factors that favoured the recent measles outbreak that occurred in Benakuma, in the North west region of Cameroon from the 21/06/2015 to 26/09/2015. We carried out a retrospective register analysis of 223 measles cases. Time trends were established using the Mann-Kendall test while survival was assessed using the Kaplan-Meier survival analysis and log rank test for comparisons. We had a case fatality of 8.5% and the mortality decreased significantly (p = 0.01) after the following public health interventions were instituted: community sensitisation and education; outbreak response immunisation and the use of a clinician in controlling the outbreak. Furthermore, the number of cases (p < 0.01) and the duration from onset of illness to consultation at a health facility (p < 0.01) decreased significantly after the institution of the aforementioned interventions. Also, survival during the outbreak was better in females (p = 0.02) and in those treated in health facilities (p < 0.01). The poor vaccination status in Benakuma coupled with negative cultural beliefs; poor socioeconomic and environmental factors and inadequate public health policies predisposed the region to a measles outbreak and favoured the spread of the virus. Public health policies should be revisited, modified and intensified to scale up vaccination coverage in measles endemic zones in Cameroon to help eliminate the disease and facilitate the overall attainment of the Sustainable Development Goals.


Clinical Outcome of Kidney Transplant Recipients with C1q-Binding De Novo Donor Specific Antibodies: A Single-Center Experience.
Complement activation by HLA antibodies is a key component of immune-mediated graft injury. We examined the clinical outcomes of kidney transplant recipients with complement-fixing de novo donor-specific antibodies (dnDSA) who were followed in our center. The C1q-binding ability was retrospectively assessed in 69 patients with dnDSA and mean fluorescence intensity (MFI) values > 2000 out of the 1325 kidney transplant recipients who were screened for DSA between 2015 and 2019. Luminex IgG single antigen beads (SAB)and C1q-SAB assays (One Lambda) were used. C1q-binding dnDSA was identified in 32/69 (46.4%) of the patients. Significantly higher MFI values were observed in C1q-positive DSA (18,978 versus 5840, p < 0.001). Renal graft biopsies were performed in 43 of the kidney transplant recipients (62.3%) with allograft dysfunction. Antibody-mediated rejection (ABMR) was detected in 29/43 (67.4%) of the patients. The incidence of ABMR was similar among patients with C1q-binding and non-C1q-binding DSA (51.7% vs. 48.3%, p = 0.523). Graft loss occurred in 30/69 (43.5%) of the patients at a median time of 82.5 months (IQR 45-135) from DSA detection. C1q-binding DSA was present in more patients who experienced graft loss (53.1% vs. 35.1%, p = 0.152). Higher MFI values and inferior clinical outcomes occurred in most of the kidney transplant recipients with C1q-binding dnDSA.


Cyclic adenosine monophosphate stimulates biosynthesis and phosphorylation of the 26 kDa gap junction protein in cultured mouse hepatocytes.
Hepatocytes prepared from 18-day-old mouse embryos were grown in serum-free medium and reached confluence after two days in culture. The total amount of the 26 kDa gap junction protein decreased in these cells during the first 24 h in culture and increased again between day 1 and day 3 more than 10-fold. At day 3 a half-life time of 2.5 to 3 h was determined for the 26 kDa protein by [35S]methionine incorporation and immunoprecipitation using affinity-purified anti-26 kDa. Incorporation of [32P]orthophosphate into the 26 kDa protein of cultured hepatocytes was found at serine residues (98%) and tyrosine residues (about 2%). The addition of dibutyryl cyclic adenosine monophosphate (db cAMP) to the culture medium at day 2 had two effects: After 15 min the extent of phosphorylation of the 26 kDa protein increased 2.7-fold whereas the total amount of the 26 kDa protein increased only 1.2-fold. After 3 h of incubation with db cAMP, a 2.5-fold increase of the 26 kDa protein was noticed which was accompanied by a 3.2-fold increase in phosphorylation of serine residues. The effects of db cAMP on phosphorylation of the 26 kDa protein could be augmented or mimicked by the addition of isoproterenol, theophylline or forskolin to the culture medium of hepatocytes. In extracts of rat hepatocarcinoma MH1C1 cells and dog kidney MDCK cells, a phosphorylated 26 kDa protein can be immunoprecipitated using anti-liver 26 kDa. These results demonstrate that the gap junction 26 kDa protein can be posttranslationally modified by cAMP-dependent phosphorylation.(ABSTRACT TRUNCATED AT 250 WORDS)


A generative model for protein contact networks.
In this paper, we present a generative model for protein contact networks (PCNs). The soundness of the proposed model is investigated by focusing primarily on mesoscopic properties elaborated from the spectra of the graph Laplacian. To complement the analysis, we also study the classical topological descriptors, such as statistics of the shortest paths and the important feature of modularity. Our experiments show that the proposed model results in a considerable improvement with respect to two suitably chosen generative mechanisms, mimicking with better approximation real PCNs in terms of diffusion properties elaborated from the normalized Laplacian spectra. However, as well as the other network models, it does not reproduce with sufficient accuracy the shortest paths structure. To compensate this drawback, we designed a second step involving a targeted edge reconfiguration process. The ensemble of reconfigured networks denotes further improvements that are statistically significant. As an important byproduct of our study, we demonstrate that modularity, a well-known property of proteins, does not entirely explain the actual network architecture characterizing PCNs. In fact, we conclude that modularity, intended as a quantification of an underlying community structure, should be considered as an emergent property of the structural organization of proteins. Interestingly, such a property is suitably optimized in PCNs together with the feature of path efficiency.


Two-Year Longitudinal Evaluation of Community Aphasia Center Participation on Linguistic, Functional Communication, and Quality of Life Measures Across People With a Range of Aphasia Presentations.
The purpose of this study was to evaluate potential changes on a hierarchy of language tasks and measures of functional communication and quality of life in a group of people with aphasia (PWA) who attended a community aphasia center for 2 years. A secondary purpose was to determine whether there were any predictors of change. Twenty-seven PWA who attended Brooks Rehabilitation Aphasia Center (BRAC) were evaluated on an aphasia battery, confrontation naming, and structured discourse in addition to completing self-reported measures of functional communication and quality of life at three time points: before attending BRAC and after 1 (N = 27) and 2 (N = 20) years of BRAC participation. Twenty-six communication partners who communicated regularly with the PWA completed a questionnaire about their functional communication at the same time points. A mixed linear model was conducted for all dependent variables to determine change over time. Tau-b correlations were conducted between demographic and aphasia-related variables and difference scores for outcome measures that exhibited significant improvements. At 1-year testing, significant improvements were observed on the aphasia battery, object and action naming, and all self- and communication partner-reported measures. At 2-year testing, all improvements were maintained except for the self-reported measure of functional communication. Structured discourse showed increases in average number of words, percentage of meaningful words and utterances, and efficiency of meaningful word production after 2 years. No significant correlations were observed between predictor variables and difference scores. Participation in aphasia centers can result in significant changes in language, functional communication, and quality of life in people with chronic aphasia. These findings support the importance of aphasia centers in the continuum of care for PWA. https://doi.org/10.23641/asha.21313689.


The hydrophobic effect: a new insight from cold denaturation and a two-state water structure.
Herein we provide a new insight into the hydrophobic effect in protein folding. Our proposition explains the molecular basis of cold denaturation, and of intermediate states in heat and their absence in cold denaturation. The exposure of non-polar surface reduces the entropy and enthalpy of the system, at low and at high temperatures. At low temperatures the favorable reduction in enthalpy overcomes the unfavorable reduction in entropy, leading to cold denaturation. At high temperatures, folding/unfolding is a two-step process: in the first, the entropy gain leads to hydrophobic collapse, in the second, the reduction in enthalpy due to protein-protein interactions leads to the native state. The different entropy and enthalpy contributions to the Gibbs energy change at each step at high, and at low, temperatures can be conveniently explained by a two-state model of the water structure. The model provides a clear view of the dominant factors in protein folding and stability. Consequently, it appears to provide a microscopic view of the hydrophobic effect and is consistently linked to macroscopic thermodynamic parameters.


The chloride/phosphate ratio combined with alkaline phosphatase as a valuable predictive marker for primary hyperparathyroidism in Chinese individuals.
The chloride/phosphate ratio (Cl/PO4) has been suggested to have a role in primary hyperparathyroidism (PHPT), but the associations between Cl/PO4 combined with ALP level and PHPT has not been well-studied. Our aim was to investigate the predictive value of combination Cl/PO4 with ALP for PHPT. A cross-sectional retrospective analysis was conducted to examine 172 patients diagnosed with PHPT categorized into two groups: normocalcaemic primary hyperparathyroidism (NPHPT) group and hypercalcaemia PHPT group. We found that Cl/PO4 levels and ALP levels in the NPHPT and hypercalcaemia PHPT group were both significantly higher than normal controls. Cl/PO4 and ALP levels were an independent risk factor for PHPT. Cl/PO4 combined with ALP increased the receiver-operating characteristic curves (ROC-AUC) and the diagnostic value in NPHPT and hypercalcaemia PHPT group (0.913; 95% CI, 0.744-1.000 and 0.932; 95% CI, 0.897-0.966, respectively), specificity of 92.8% and sensitivity of 98%. In conclusion, combination Cl/PO4 with ALP might be a low-cost, simple, available predictive marker of PHPT in Chinese individuals, particularly Chinese remote region where the method used to measure PTH cannot be done. Moreover, due to serum calcium level in NPHPT, Cl/PO4 combined with ALP level measurement have great potential to predict significant occurrence of NPHPT.


Brain-derived neurotrophic factor mechanisms and function in adult synaptic plasticity: new insights and implications for therapy.
Synaptic plasticity, the fiundamental ability of synapses to undergo experience-dependent changes in synaptic strength, is thought to underlie a range of long-term adaptive responses of the central nervous system. The inability to generate or maintain appropriate changes in synaptic connectivity and neuronal circuitry is likely to impair cognitive futnction associated with many neurological and psychiatric disorders. The success of intervening therapeutic approaches will depend on the ability to target key component mechanisms in the complex process of synaptic plasticity. Brain-derived neurotrophic factor (BDNF) has emerged as a major regulator of synaptic plasticity and a key target in disorders such as major depression and Alzheimner's disease. This review discusses findings that link BDNF mechanisms in synaptic plasticity to disease mechanisms and possible future therapies.


Cu(2)ZnSiS(4).
Single crystals of Cu(2)ZnSiS(4), dicopper(I) zinc silicon tetrasulfide, have been prepared via high-temperature solid-state synthesis. Cu(2)ZnSiS(4) was found to have the wurtz-stannite structure type, like that of Li(2)CdGeS(4), Li(2)CdSnS(4), and Cu(2)CdSiS(4). Each sulfur anion is tetra-hedrally coordinated by two Cu cations, one Si cation, and one Zn cation, forming a three-dimensional honeycomb structure. When viewed along the c axis, the atoms are aligned in rows in which each cation alternates with the sulfur anions.


The Typical Presentation Spectrum of Deep Vein Thrombosis Associated with Inferior Vena Cava Malformations.
Congenital malformations of the inferior vena cava (IVC) are rare and underreported. They can be a risk factor for deep venous thrombosis (DVT) as a result of inadequate venous drainage of the lower extremities through collateral circulation. The significant number of cases reported in the literature highlights their importance, warranting investigating their existence in younger individuals with idiopathic DVT of the lower extremities and pelvic veins. In this systematic review, we depict the typical presentation of IVC malformations, their management, and the management of their associated DVT.


[Plea for the retention of the Dutch National Medical Registration (LMR) to provide reliable information regarding public health and healthcare].
Since 1963, information on each hospital admission has been collected from all general and university hospitals in the Netherlands (the 'Landelijke Medische Registratie'; LMR). This information includes, among others, one main discharge diagnosis and up to nine secondary diagnoses made during admission, the duration of the hospital stay, death during admission, times of admission and discharge, type of specialist who treated the patient, operations and related therapeutic procedures. This information system is not biased by financial incentives and is therefore a good resource for epidemiological studies and drug-safety research. As of 2005, a new type of administration has been introduced that records diagnosis-therapy combinations for reimbursement purposes. Because the reimbursement of closely related diagnosis-therapy combinations may differ, the system is not only sensitive to fraud but will probably also jeopardise the validity of epidemiological research with such data. Therefore, the LMR system cannot be missed.


Integration of sensory information via central thermoregulatory leptin targets.
The adipocyte derived hormone leptin acts in the brain to regulate body weight, food intake and energy expenditure. Even though it is well accepted that leptin regulates energy expenditure at least in part by modulating thermogenesis, the exact mechanisms are not clear. Particularly, it is unclear which central circuits regulate thermogenic leptin actions and if and how these may interact with feeding circuits. Within the last decade our understanding of central thermoregulatory circuits has increased substantially and allowed the identification of leptin target neurons (those expressing the long form leptin receptor - LepRb) that are involved in the sympathetic control of the heat generating brown adipose tissue (BAT). Indeed, LepRb neurons in the preoptic area and dorsomedial hypothalamus are part of the known thermoregulatory circuits controlling sympathetic premotor neurons that are located in the raphe pallidus. Thermoregulatory control and food intake are both regulated by leptin signaling pathways, even though distinct neuronal pathways have been described, respectively. Nevertheless, feeding status and control of body temperature and energy expenditure are tightly interconnected, but it is unknown how these aspects are connected within leptin signaling pathways to result in appropriate output signals (e.g. BAT thermogenesis). Indeed, cold-induced thermogenesis is potently blocked during fasting, which instead triggers an active decrease in energy expenditure and body temperature, a state known as torpor. In this article we will review recent data characterizing central thermoregulatory LepRb pathways and speculate on potential integration mechanisms that may relay anorexic and thermoregulatory leptin action to control energy homeostasis.


Studies on gastrointestinal tract functional changes in diabetic animals.
Diarrhea and constipation are frequently reported gastrointestinal complications in diabetic patients. These disorders seem to be the consequence of altered innervations of a receptor system in the gastrointestinal tract in diabetes. We investigated the functional changes of cholinergic and beta-adrenergic receptor activities in rat ileum tissue after 8 weeks of control and streptozotocin (STZ)-induced diabetes in rats. Functional changes with different agonists were observed in diabetes: In diabetic rat ileum, carbachol increased the maximal contraction without changing pD2 values, whereas with acetylcholine, no changes were observed in the maximal contractile (Emax) response and pD2 values in diabetes were comparable with controls. With the beta-adrenoceptor agonists isoproterenol and salmeterol, decreased maximal relaxation responses were observed in 8-week diabetic rat ileums as compared with age-matched controls. With isoproterenol, pD2 values were also decreased in diabetic ileum, although not with salmeterol. On the other hand, with the in vivo charcoal meal test, the percentage of transit was increased in diabetic mice. The antitransit percentage was significantly increased in diabetic mice with the muscarinic antagonist atropine and the beta 2-selective agonist salbutamol, as compared with age-matched control mice. Results obtained from these in vivo and in vitro studies indicate that gastrointestinal complications such as diabetic diarrhea may occur because of functional changes such as increased intestinal transit and decreased intestinal tone due to increased cholinergic and decreased beta-adrenergic receptor activities in diabetes mellitus.


Precautions When Providing Dental Care During Coronavirus Disease 2019 (COVID-19) Pandemic.
Transmission of coronavirus disease 2019 (COVID-19)-caused by novel severe acute respiratory syndrome coronavirus 2-through aerosolised saliva and respiratory droplets is possible when aerosol-generating dental procedures are performed. Consequently, dental practitioners are at increased risk of being infected when treating COVID-19 patients. A comprehensive review of the current literature on precautions when providing dental care during the COVID-19 pandemic is discussed and recommendations for dental practitioners are made. Dental practitioners should actively keep themselves abreast of the guidelines published by both national and international authorities and adhere strictly to them.


Kainate-induced delayed onset of excitotoxicity with functional loss unrelated to the extent of neuronal damage in the in vitro spinal cord.
While excitotoxicity is a major contributor to the pathophysiology of acute spinal injury, its time course and the extent of cell damage in relation to locomotor network activity remain unclear. We used two in vitro models, that is, the rat isolated spinal cord and spinal organotypic cultures, to explore the basic characteristics of excitotoxicity caused by transient application of the glutamate analogue kainate followed by washout and analysis 24 h later. Electrophysiological records showed that fictive locomotion was slowed down by 10 microM kainate (with no histological loss) and fully abolished by 50 microM, while disinhibited bursting with unchanged periodicity persisted. Kainate concentrations (> or =50 microM) larger than those necessary to irreversible suppress fictive locomotion could still elicit dose-dependent motoneuron pool depolarization, and dose-dependent neuronal loss in the grey matter, especially evident in central and dorsal areas. Motoneuron numbers were largely decreased. A similar regional pattern was detected in organotypic slices, as extensive cell loss was dose related and affected motoneurons and premotoneurons: the number of dead neurons (already apparent 1 h after kainate) grew faster with the higher kainate concentration. The histological damage was accompanied by decreased MTT formazan production commensurate with the number of surviving cells. Our data suggest locomotor network function was very sensitive to excitotoxicity, even without observing extensive cell death. Excitotoxicity developed gradually leaving a time window in which neuroprotection might be attempted to preserve circuits still capable of expressing basic rhythmogenesis and reconfigure their function in terms of locomotor output.


Membrane-integrated microfluidic device for high-resolution live cell imaging.
The design and fabrication of a membrane-integrated microfluidic cell culture device (five layers,≤500 μm total thickness) developed for high resolution microscopy is reported here. The multi-layer device was constructed to enable membrane separated cell culture for tissue mimetic in vitro model applications and pharmacodynamic evaluation studies. The microdevice was developed via a unique combination of low profile fluidic interconnect design, substrate transfer methodology, and wet silane bonding. To demonstrate the unique high resolution imaging capability of this device, we used oil immersion microscopy to image stained nuclei and mitochondria in primary hepatocytes adhered to the incorporated membrane.


Different voltage dependence of transient and persistent Na+ currents is compatible with modal-gating hypothesis for sodium channels.
1. These experiments tested the hypothesis that the differing voltage dependence of the transient (INa) and persistent (INaP) Na+ currents in neocortical neurons results from the state of inactivation of one type of Na+ channel rather than from the existence of different types of Na+ channels. This question was examined in acutely isolated pyramidal neurons from the sensorimotor cortex of rats by using papain to remove inactivation from INa and comparing the resulting activation curve with that of INaP. 2. In control cells, INaP activated at more negative potentials than INa. Inclusion of papain in the recording pipette removed inactivation from INa and caused the INa activation curve to be shifted leftward to the position of the curve for INaP measured in control cells. Papain greatly increased both INa amplitude and the time to reach peak INa during smaller depolarizations, whereas the difference between control and test currents was reduced during large depolarizations. 3. We conclude that differences in the voltage dependence of INa and INaP activation does not provide sufficient evidence that these currents flow through separate sets of Na+ channels. Instead, our results are consistent with the idea that INaP largely arises from a fraction of the transient Na+ channels that intermittently lose their inactivation.


Preparation and characterization of liposome-encapsulated haemoglobin by a freeze-thaw method.
Haemoglobin (Hb) was encapsulated into liposomes as a blood substitute by a freeze-thaw method. The encapsulation efficiency was affected by the Hb/lipid ratio, starting Hb concentration, pH and salt concentration. Liposome-encapsulated haemoglobin (LEH) prepared by this method contains 5-10 mM Hb with 4-10 per cent methaemoglobin (Met-Hb), depending on the starting Hb/lipid ratio and Met-Hb content. The encapsulated Hb has the same absorption spectrum as free Hb and shows oxygen-dissociation characteristics similar to normal red blood cells when 2,3-diphosphoglycerate is co-entrapped in the liposomes. LEH exhibited some leakage, which was greatly reduced by sequential extrusions of LEH through polycarbonate membranes (1.0 and 0.45 microns). Stability of LEH was studied using different Hb preparations, and antioxidants of lipids or/and Hb either at 4 or 37 degrees C. alpha-tocopherol or butylated hydroxytoluene, antioxidants of lipids, inhibited not only the peroxidation of liposomes but also Hb oxidation. Among antioxidants of Hb, NADH was most effective in preventing the oxidation of Hb. Glutathione had a moderate preventive effect. However, catalase had no effect and ascorbate accelerated the oxidation of Hb. Glucose and glutathione decreased the oxidation of Hb only in the Hb preparation obtained by hypotonic lysing, not in that by toluene lysing. These results indicate that the Met-Hb reductase system in the latter is lost or inactivated during isolation.


Meiotic linkage mapping of 52 genes onto the canine map does not identify significant levels of microrearrangement.
In an effort to extend our understanding of the evolutionary relationship between the canine and human genomes, we have developed and positioned 52 new gene-associated polymorphic markers on the canine meiotic linkage map. Canine-specific PCR primers were developed from the consensus of published sequences of several mammalian genomes and were designed to span intronic regions, thus optimizing the probability that a polymorphic site was included. The resulting markers were analyzed on a panel of three-generation canine reference families and the data were incorporated into the current meiotic linkage map. The data were compared with those generated by three chromosome paint studies in an effort to understand the distribution and frequency of microrearrangements within the canine genome. Forty-eight of 52 genes map to a chromosomal region predicted to contain genes from the corresponding region of the human genome according to all published reciprocal chromosome paint studies. Meiotic linkage mapping data for three genes can be used to resolve discrepancies between the published reciprocal chromosome paint studies, and for an additional two genes, meiotic mapping data allow evolutionary breakpoints to be more precisely defined. We conclude that microrearrangements of evolutionarily conserved segments between the canine and human genomes are rare, occurring for less than 0.5% of gene data reported to date. In addition, we have found that the placement of genes on the meiotic linkage map is a useful mechanism for resolving discrepancies between existing data sets.


[Extraskeletal manifestations of seronegative spondyloarthropathies].
Spondyloarthropathies constitute a cluster of interrelated and overlapping chronic inflammatory rheumatic diseases of unknown etiology that include ankylosing spondylitis, psoriatic arthritis, reactive arthritis, enteropathic arthritis and undifferentiated spondyloarthritis. These diseases are not associated with rheumatoid factor, they occur in genetically predisposed individuals and show a strong association with HLA-B27. The primary pathologic sites include the entheses, the sacroiliac joints and the axial skeleton with or without some nonarticular structures, such as the skin, eye, aortic valve, heart and kidney.


Imputing missing genotypes with weighted k nearest neighbors.
Missing values are a common problem in genetic association studies concerned with single-nucleotide polymorphisms (SNPs). Since many statistical methods cannot handle missing values, such values need to be removed prior to the actual analysis. Considering only complete observations, however, often leads to an immense loss of information. Therefore, procedures are required that can be used to impute such missing values. In this study, an imputation procedure based on a weighted k nearest neighbors algorithm is presented. This approach, called KNNcatImpute, searches for the k SNPs that are most similar to the SNP whose missing values need to be replaced and uses these k SNPs to impute the missing values. Alternatively, KNNcatImpute can search for the k nearest subjects. In this situation, the missing values of an individual are imputed by considering subjects showing a DNA pattern similar to the one of this individual. In a comparison to other imputation approaches, KNNcatImpute shows the lowest rates of falsely imputed genotypes when applied to the SNP data from the GENICA study, a candidate SNP study dedicated to the identification of genetic and gene-environment interactions associated with sporadic breast cancer. Moreover, KNNcatImpute can also be applied to data from genome-wide association studies, as an application to a subset of the HapMap data demonstrates.


Construction of chicken x human microcell hybrids for human gene targeting.
Human chromosomes 1, 2, 3, and 11 were tagged with pSV2 neo and transferred via microcell fusion from mouse A9 human monochromosomal hybrids to a chicken pre-B cell line, DT40, proficient for homologous recombination. Hybrids containing two copies of human chromosome 11 were transfected with targeting vectors containing a mammalian selectable gene with either the D11S16 or HRAS genomic sequences corresponding to two different chromosome 11 loci. Analysis of stable transfectants showed a high frequency (approximately 80%) of targeted integration of these constructs into each of the homologous loci of human chromosome 11 in DT40 hybrids. The results suggest that any human genomic sequences on human chromosomes transferred into DT40 cells could be targeted at high frequency, thereby allowing for subsequent modification of human genes and chromosomes.


The role of tinman, a mesodermal cell fate gene, in axon pathfinding during the development of the transverse nerve in Drosophila.
During the development of peripheral nerves, pioneer axons often navigate over mesodermal tissues. In this paper, we examine the role of the mesodermal cell determination gene tinman on cells that provide pathfinding cues in Drosophila. We focus on a subset of peripheral nerves, the transverse nerves, that innervate abdominal segments. During wildtype embryonic development, the transverse nerve efferents associate with glial cells located on the dorsal aspect of the CNS midline (transverse nerve exit glia). These glial cells have cytoplasmic extensions that prefigure the transverse nerve pathway from the CNS to the body wall musculature prior to transverse nerve formation. Transverse nerve efferents extend along this scaffold to the periphery, where they fasciculate with projections from a peripheral neuron--the LBD. In tinman mutants, the transverse nerve exit glia appear to be missing, and efferent fibers remain stalled at the CNS midline, without forming transverse nerves. In addition, fibers of the LBD neurons are often truncated. These results suggest that the lack of exit glia prevents normal transverse nerve pathfinding. Another prominent defect in tinman is the loss of all dorsal neurohemal organs, FMRFamide-expressing thoracic structures which likely contain the homologs of the transverse nerve exit glia in the thoracic segments. Our results support the hypothesis that the exit glia have a mesodermal origin and that glia play an essential role in determining transverse nerve axon pathways.


Common gastrointestinal drug-drug interactions in geriatrics and the importance of careful planning.
Polypharmacy, which uses multiple medications to treat chronic illnesses, is common among elderly patients. However, it can lead to drug interactions, especially with gastrointestinal (GI) medicines that are extensively used. These drug interactions can have severe consequences and pose a significant challenge to healthcare providers. Therefore, it is crucial to identify the underlying mechanisms of these interactions and develop strategies to minimize medication errors. We analyzed databases on GI illnesses common in older adults, including GERD, peptic ulcer disease, IBS, IBD, constipation, and diarrhea. Our research identified noteworthy drug interactions and utilized major electronic databases such as USFDA, PubMed, Scopus, and Google Scholar until 15 May 202315 May 2023, along with a review of reference lists. Aging can affect how the body processes drugs, leading to an increased risk of drug interactions. Therefore, healthcare professionals must carefully evaluate a patient's medical history and health condition to design personalized treatment plans.


Tuberous Sclerosis Complex Associated Neuropsychiatric Disorders and Parental Stress: Findings from a National, Prospective TSC Surveillance Study.
To assess the role of the TAND (tuberous sclerosis complex [TSC] associated neuropsychiatric disorders) checklist as a screening tool for neuropsychiatric pathology, to evaluate behavioral and psychiatric symptoms and related parental stress in children with TSC, and to analyze associations between parental stress, TAND findings, and TSC pathology. This is a prospective cohort study including 22 individuals from a national TSC surveillance study in Germany using demographic and clinical data, the TAND checklist, the Child Behavior Checklist (CBCL), and the Parenting Stress Index (PSI). Mean (standard deviation) age at follow-up was 4 years (3 years 9 months), and 13/22 of patients were male. Seventeen children had epilepsy (focal: 9; generalized: 4; infantile spasms: 4). Developmental delay was diagnosed in 12/22 patients. The most prevalent TAND items were anxiety and mood swings in 10/22 children. At least one TAND item was reported by 17/22 patients, internalizing symptoms by 10/22, and externalizing symptoms by 11/22. In contrast, only one patient had a clinically relevant score in the CBCL scales. Of 22 parents, 12 reported clinically relevant parental stress due to both child and parenting factors. Higher total parental stress was associated with a higher TAND externalizing score (r = 0.49; p = 0.028) and TAND total score (r = 0.51; p = 0.016), a higher CBCL total score (r = 0.59; p = 0.005), and the number of antiepileptic drugs (r = 0.50; p = 0.017). Developmental delay was correlated with child stress factors (r = 0.48; p = 0.023). The TAND checklist appears to be a promising screening tool for neuropsychiatric problems in very young children with TSC. Parental stress in children with TSC is modified by TSC-related pathology, both neuropsychiatric and neurological.


In Vitro Activity of Tedizolid against Mycobacterium tuberculosis.
Tedizolid is a novel oxazolidinone with activities against Gram-positive microorganisms, including mycobacteria. We studied the in vitro activity of tedizolid against 120 Mycobacterium tuberculosis strains, including susceptible, first-line-resistant, and multidrug-resistant isolates. MIC was tested using the Bactec 960 MGIT system. MIC90 and MIC50 were 0.5 and 0.25 μg/ml, respectively, in susceptible and resistant strains. Tedizolid may be an alternative in the treatment of resistant M. tuberculosis.


Improvements of mean body mass index and body weight in preobese and overweight Japanese adults with black Chinese tea (Pu-Erh) water extract.
Water-soluble black Chinese (Pu-Erh) tea extract (BTE), which contains high gallic acid content, has been demonstrated to elicit antiobese effects in animals. Because gallic acid is related with the reduction of visceral fat and cholesterol contents and improvement of obesity in animals, we investigated the effects of BTE intake on 36 preobese Japanese adults (body mass index [BMI], >25- <30 kg/m(2)) in a 12-week double-blind, randomized, placebo-controlled group comparison study using powdered barley tea with or without (placebo) BTE. A follow-up 4-week period after BTE intake termination was monitored to observe the withdrawal effect. All subjects ingested barley tea with or without BTE (333 mg) before each of the 3 daily meals. In the BTE-treated group, the mean pretreament values of body weight and BMI significantly decreased after intake and after BTE withdrawal. However, the corresponding values scored significant differences only from 8 weeks after intake (vs the placebo-treated group). The mean values of the waist circumference indicated a similar tendency. Furthermore, coronal navel section (same anatomical position) images of computed tomography of all BTE- and non-BTE-treated subjects revealed that the visceral fat areas (cm(2)) were significantly (P < .05) less in the former 12 weeks after BTE ingestion. Measured biochemical parameters did not indicate significant differences, and BTE-treated subjects did not complain of any adverse effects (abdominal distension, etc). Ingestion of BTE exhibited significant effects in reducing the mean waist circumference, BMI, and visceral fat values and might be useful for weight control and prevention of obesity development (or metabolic syndrome) in humans.


Evaluation of the initial implementation of a nationwide diabetic retinopathy screening programme in primary care: a multimethod study.
The Australian Government funded a nationwide diabetic retinopathy screening programme to improve visual outcomes for people with diabetes. This study examined the benefits and barriers of the programme, image interpretation pathways and assessed the characteristics of people who had their fundus photos graded by a telereading service which was available as a part of the programme. Multimethod: survey and retrospective review of referral forms. Twenty-two primary healthcare facilities from urban, regional, rural and remote areas of Australia, and one telereading service operated by a referral-only eye clinic in metropolitan Sydney, Australia. Twenty-seven primary healthcare workers out of 110 contacted completed a survey, and 145 patient referrals were reviewed. Manifest qualitative content analysis showed that primary healthcare workers reported that the benefits of the screening programme included improved patient outcomes and increased awareness and knowledge of diabetic retinopathy. Barriers related to staffing issues and limited referral pathways. Image grading was performed by a variety of primary healthcare workers, with one responder indicating the utilisation of a diabetic retinopathy reading service. Of the people with fundus photos graded by the reading service, 26.2% were reported to have diabetes. Overall, 12.3% of eyes were diagnosed with diabetic retinopathy. Photo quality was rated as excellent in 46.2% of photos. Referral to an optometrist for diabetic retinopathy was recommended in 4.1% of cases, and to an ophthalmologist in 6.9% of cases. This nationwide diabetic retinopathy screening programme was perceived to increase access to diabetic retinopathy screening in regional, rural and remote areas of Australia. The telereading service has diagnosed diabetic retinopathy and other ocular pathologies in images it has received. Key barriers, such as access to ophthalmologists and optometrists, must be overcome to improve visual outcomes.


Women doctors for women patients?
This review concentrates on two main areas: (i) the attitudes of women patients towards physicians and (ii) the attitudes of physicians towards female patients. Women are important users of the health care services. There is female to male ratio of 3.8:3.0 for the average number of consultations in general practice per person per year. Women are prescribed more drugs than men; in 1974 66 per cent of women and 54 per cent of men had at least one drug prescribed. Physician gender is thought to have an effect through the physician-patient relationship and its outcomes. Patients perceive male and female doctors differently. Women patients believe women doctors to have the good qualities of both male and female physicians, e.g. assertiveness and initiative but also tenderness and nurturance. There is some evidence that women doctors give their female patients more surgery time than is allocated to them by male doctors. In the specialty areas, preference for a female gynaecologist is stated more often than a preference for a female doctor in any other specialty. The attitudes of physicians of both sexes towards female patients is seen by some as negative; however, physicians as a group are more 'feminist' than control groups. Some women GPs are seeing up to 85 per cent female patients.


Characterizing sympathetic neurovascular transduction in humans.
Despite its critical role for cardiovascular homeostasis in humans, only a few studies have directly probed the transduction of sympathetic nerve activity to regional vascular responses--sympathetic neurovascular transduction. Those that have variably relied on either vascular resistance or vascular conductance to quantify the responses. However, it remains unclear which approach would better reflect the physiology. We assessed the utility of both of these as well as an alternative approach in 21 healthy men. We recorded arterial pressure (Finapres), peroneal sympathetic nerve activity (microneurography), and popliteal blood flow (Doppler) during isometric handgrip exercise to fatigue. We quantified and compared transduction via the relation of sympathetic activity to resistance and to conductance and via an adaptation of Poiseuille's relation including pressure, sympathetic activity, and flow. The average relationship between sympathetic activity and resistance (or conductance) was good when assessed over 30-second averages (mean R(2) = 0.49±0.07) but lesser when incorporating beat-by-beat time lags (R(2) = 0.37±0.06). However, in a third of the subjects, these relations provided relatively weak estimates (R(2)<0.33). In contrast, the Poiseuille relation reflected vascular responses more accurately (R(2) = 0.77±0.03, >0.50 in 20 of 21 individuals), and provided reproducible estimates of transduction. The gain derived from the relation of resistance (but not conductance) was inversely related to transduction (R(2) = 0.37, p<0.05), but with a proportional bias. Thus, vascular resistance and conductance may not always be reliable surrogates for regional sympathetic neurovascular transduction, and assessment from a Poiseuille relation between pressure, sympathetic nerve activity, and flow may provide a better foundation to further explore differences in transduction in humans.


Peptide linkage mapping of the Agrobacterium tumefaciens vir-encoded type IV secretion system reveals protein subassemblies.
Numerous bacterial pathogens use type IV secretion systems (T4SS) to deliver virulence factors directly to the cytoplasm of plant, animal, and human host cells. Here, evidence for interactions among components of the Agrobacterium tumefaciens vir-encoded T4SS is presented. The results derive from a high-resolution yeast two-hybrid assay, in which a library of small peptide domains of T4SS components was screened for interactions. The use of small peptides overcomes problems associated with assaying for interactions involving membrane-associated proteins. We established interactions between VirB11 (an inner membrane pore-forming protein), VirB9 (a periplasmic protein), and VirB7 (an outer membrane-associated lipoprotein and putative pilus component). We provide evidence for an interaction pathway, among conserved members of a T4SS, spanning the A. tumefaciens envelope and including a potential pore protein. In addition, we have determined interactions between VirB1 (a lytic transglycosylase likely involved in the local remodeling of the peptidoglycan) and primarily VirB8, but also VirB4, VirB10, and VirB11 (proteins likely to assemble the core structure of the T4SS). VirB4 interacts with VirB8, VirB10, and VirB11, also establishing a connection to the core components. The identification of these interactions suggests a model for assembly of the T4SS.


Vaccination coverage and associated factors for receipt of the 23-valent pneumococcal polysaccharide vaccine in Taiwan: A nation-wide community-based study.
Older adults, particularly those with chronic obstructive pulmonary disease, are advised to receive 23-valent pneumococcal polysaccharide vaccine (PPV23). However, the PPV23 vaccination rate in Taiwan and the determinants of receipt remain unclear.We analyzed the data of 1475 community-dwelling older adults aged ≥75 years who participated in the Healthy Aging Longitudinal Study in Taiwan. Each participant received assessments of PPV23 status, sociodemographic factors (age, sex, education level, marital status, living alone, and occupation), and health-related factors (chronic diseases, smoking status, alcohol intake, physical activities, cognitive status, and physical performance). PPV23 rate was defined as the number of participants who reported receiving free PPV23 divided by the total number of candidates for free PPV23. Multinomial logistic regression analysis was applied to investigate the sociodemographic and health-related determinants of PPV23 status.A PPV23 vaccination rate of 20.7% (305/1475) was observed. Participants who were female, current smokers, and had a low peak expiratory flow were associated with PPV23 nonreceipt (all P <.05). Of the participants who had a low peak expiratory flow, low education status, and physical inactivity were associated with PPV23 nonreceipt (all P <.05).The PPV23 vaccination rate among adults aged ≥75 years was low. Older adults who were women, current smokers, or who had a low PEF were less likely to receive the PPV23. These findings support continual efforts to improve the PPV23 coverage rate in vulnerable populations.


Bleomycin alone or combined with mitomycin C in treatment of advanced or recurrent squamous cell carcinoma of the vulva.
Twenty patients with advanced and/or recurrent vulvar carcinoma were treated with bleomycin alone or combined with mitomycin C. Five of 11 patients in the bleomycin-treated group showed objective response (two complete and three partial) and five of nine patients in the bleomycin-mitomycin C-treated group showed objective response (one complete and four partial). No severe side effects were seen. Now that activity against this disease has been demonstrated for bleomycin, alone or combined with mitomycin C, attention should be directed to treating those patients not cured by surgery and radiation therapy alone.


Gonadotrope-specific expression and regulation of ovine follicle stimulating hormone Beta: transgenic and adenoviral approaches using primary murine gonadotropes.
The beta subunit of follicle stimulating hormone (FSHB) is expressed specifically in pituitary gonadotropes in vertebrates. Transgenic mouse studies have shown that enhancers in the proximal promoter between -172/-1 bp of the ovine FSHB gene are required for gonadotrope expression of ovine FSHB. These enhancers are associated with regulation by activins and gonadotropin releasing hormone (GnRH). Additional distal promoter sequence between -4741/-750 bp is also required for expression. New transgenic studies presented here focus on this distal region and narrow it to 1116 bp between -1866/-750 bp. In addition, adenoviral constructs were produced to identify these critical distal sequences using purified primary mouse gonadotropes as an in vitro model system. The adenoviral constructs contained -2871 bp, -750 bp or -232 bp of the ovine FSHB promoter. They all showed gonadotrope-specific regulation since they were induced only in purified primary gonadotropes by activin A (50 ng/ml) and inhibited by GnRH (100 nM) in the presence of activin (except -232FSHBLuc). However, basal expression of all three viral constructs (in the presence of follistatin to block cellular induction by activin) was relatively high in pituitary non-gonadotropes as well as gonadotropes. Thus, gonadotrope-specific regulation associated with the proximal promoter was observed as expected, but the model was blind to distal promoter elements between -2871/-750 necessary for gonadotrope-specific expression of ovine FSHB in vivo. The new adenoviral-based in vitro technique did detect, however, a novel GnRH response element between -750 bp and -232 bp of the ovine FSHB promoter. We conclude that adenoviral-based studies in primary gonadotropes can adequately recognize regulatory elements on the ovine FSHB promoter associated with gonadotrope-specific regulation/expression, but that more physiologically based techniques, such as transgenic studies, will be needed to identify sequences between -1866/-750 bp of the ovine FSHB promoter that are also required for tissue/cell specific expression in vivo.


Microbiota and Human Reproduction: The Case of Female Infertility.
During the last decade, the availability of next-generation sequencing-based approaches has revealed the presence of microbial communities in almost all the human body, including the reproductive tract. As for other body sites, this resident microbiota has been involved in the maintenance of a healthy status. As a consequence, alterations due to internal or external factors may lead to microbial dysbiosis and to the development of pathologies. Female reproductive microbiota has also been suggested to affect infertility, and it may play a key role in the success of assisted reproductive technologies, such as embryo implantation and pregnancy care. While the vaginal microbiota is well described, the uterine microbiota is underexplored. This could be due to technical issues, as the uterus is a low biomass environment. Here, we review the state of the art regarding the role of the female reproductive system microbiota in women's health and human reproduction, highlighting its contribution to infertility.


Enhanced release of prostaglandin D2 during re-incubation of RAW 264.7 macrophage-like cells after treatment of both lipopolysaccharide and non-steroidal anti-inflammatory drugs.
RAW 264.7 macrophage-like cells are known to release prostaglandins (PGs), mainly PGD(2) to the culture medium after lipopolysaccharide (LPS)-treatment. This release was inhibited by non-steroidal anti-inflammatory drugs (NSAIDs), which are known to inhibit prostaglandin H(2) synthase (PGHS) activity. In this study, we examined the effect of removal of NSAID after induction of PGHS with LPS, on the release of PGs, which has not been studied well. Re-incubation of RAW 264.7 cells after treatment of both LPS and NSAIDs resulted in enhanced release of PGD(2) compared with the cells pretreated with LPS alone. Besides, PGHS activity was detectable in these cell homogenates and the amount of PGHS-2 protein showed similar changes to PGD(2) release. However, addition of NSAIDs again in the re-incubation period almost completely inhibited the PGD(2) release but increased the amount of PGHS-2 protein to the higher levels. Various types of NSAIDs used in this study showed similar effects on the changes in PGD(2) release and PGHS-2 protein amounts, except those on PGHS activity in cell homogenates; while indomethacin, aspirin, and NS-398 inhibited it, but nimesulide and acetaminophen did not. These results seem to suggest an importance for the caution that the enhanced induction of PGHS-2 protein and the concomitant release of PGs release would occur after removal of the NSAID not only from the medium in in vitro experiments but also from therapeutic prescription.


Refluxing supernumerary kidney: easy to overlook.
Supernumerary kidney is a rare anomaly and most of the times it is incidentally detected. On occasions it may present with symptoms due to stones, tumours and infections. Supernumerary unit if small and dysplastic may easily escape detection. In this case report, supernumerary kidney presented as vesicoureteric reflux in a 4-year-old male child.


Behavioral and social sciences and public health at CDC.
Although the history of CDC spans 60 years, only during the last 2 decades of the 20th century did the agency come to recognize and better understand the importance of the behavioral and social sciences to its overall mission. This recognition was a consequence of several events, notably the growing public awareness of the many conditions and diseases linked to unhealthy behavior and the creation of three new organizational units at CDC--the National Center for Chronic Disease Prevention and Health Promotion in 1988, the National Center for Injury Prevention and Control in 1992, and the National Center for HIV, STD, and TB Prevention in 1993--that focused on conditions, diseases, and injuries with clear behavioral risks. Accordingly, the relatively small number of CDC behavioral and social scientists were initially concentrated in these three centers. In 1995, to raise awareness of behavioral and social sciences at CDC and to integrate these fields into CDC-conducted and -supported research and practice activities, the agency's behavioral and social scientists established the Behavioral and Social Sciences Working Group (BSSWG). The application of the behavioral and social science disciplines to public health attests to the success of the working group. Today, BSSWG continues as a formal organization sponsored by the Office of the Chief Science Officer within the CDC Office of the Director.


First case of feline systemic Cryptococcus albidus infection.
This paper, as best as the authors can determine, is the first to describe a documented case of systemic infection caused by Cryptococcus albidus in a cat. The patient had a history of paralysis of the hind legs and had been treated with prednisone for 1 month. Microscopic examination of a fine needle biopsy specimen from a right popliteal lymph node showed granulomatous inflammation with many encapsulated yeast cells. Moreover, microscopic examination of Indian ink preparations of the cerebrospinal fluid revealed encapsulated ovoid yeast cells. Thus this case was diagnosed to be cryptococcosis. However, the cat died after treatment for three days with voriconazole. Isolates recovered from samples of the cerebrospinal fluid, liver and spleen were identified as C. albidus by molecular analysis, as well as through morphologic and biochemical studies. Therefore, this case indicates that C. albidus should be considered as a potential feline pathogen.


Tuning of Thermoresponsivity of a Poly(2-alkyl-2-oxazoline) Block Copolymer by Interaction with Surface-Active and Chaotropic Metallacarborane Anion.
Thermoresponsive nanoparticles based on the interaction of metallacarboranes, bulky chaotropic and surface-active anions, and poly(2-alkyl-2-oxazoline) block copolymers were prepared. Recently, the great potential of metallacarboranes have been recognized in biomedicine and many delivery nanosystems have been proposed. However, none of them are thermoresponsive. Therefore, a thermoresponsive block copolymer, poly(2-methyl-2-oxazoline)-block-poly(2-n-propyl-2-oxazoline) (PMeOx-PPrOx), was synthesized to encapsulate metallacarboranes. Light scattering, NMR spectroscopy, isothermal titration calorimetry, and cryogenic TEM were used to characterize all solutions of the formed nanoparticles. The cloud-point temperature (TCP ) of the block copolymer was observed at 30 °C and polymeric micelles formed above this temperature. Cobalt bis(dicarbollide) anion (COSAN) interacts with both polymeric segments. Depending on the COSAN concentration, this affinity influenced the phase transition of the thermoresponsive PPrOx block. The TCP shifted to lower values at a lower COSAN content. At higher COSAN concentrations, the hybrid nanoparticles are fragmented into relatively small pieces. This system is also thermoresponsive, whereby an increase in temperature leads to higher polymer mobility and COSAN release.


The effect of cigarette smoking on the detection of small radiographic opacities in inorganic dust diseases.
Whether cigarette smoking can cause radiographic opacities indistinguishable from those due to pneumoconiosis remains controversial. The situation becomes clearer when one limits the abnormalities to those that can be standardized under the International Labour Office (ILO) classification system. The bulk of the evidence indicates that, using the ILO system, cigarette smoking alone is not associated with radiographic opacities that would be mistaken for pneumoconiosis with sufficient frequency to be of any practical importance. The effects of cigarette smoking, as a cofactor, in conjunction with occupational dust exposure depend on the type of dust. No relationship has been convincingly demonstrated for coal dust or silica. Only with asbestos exposure does there appear to be a significant cigarette smoking-associated increase in the frequency of irregular radiographic opacities. This increase does not appear to translate into a restrictive impairment in pulmonary function. The limited information available indicates that the features of asbestosis on high-resolution computed tomography are not similarly related to cigarette smoking. Additional research is needed to substantiate the relationship between smoking and occupational exposure to dust of many types, and also the possible imaging and pathophysiologic significance of their interactions.


[Deficiency of Magnesium and Pyridoxine as a Risk Factors for Coronary Heart Disease].
Deficiency of magnesium and vitamin 6 is a modifiable risk factor for coronary heart disease (CHD). A combined deficiency of magnesium and vitamin 6 disrupts the functioning myocardial tissue due to the formation of endothelial dysfunction, prothrombotic changes in blood viscosity characteristics, improve vascular tone, failure of the mitochondria. Receiving organic magnesium salts in combination with adequate doses of pyridoxine can be effective in the prevention and treatment of CHD.


Reduced explicitly correlated Hartree-Fock approach within the nuclear-electronic orbital framework: applications to positronic molecular systems.
In the application of the nuclear-electronic orbital (NEO) method to positronic systems, all electrons and the positron are treated quantum mechanically on the same level. Explicit electron-positron correlation can be included using Gaussian-type geminal functions within the variational self-consistent-field procedure. In this paper, we apply the recently developed reduced explicitly correlated Hartree-Fock (RXCHF) approach to positronic molecular systems. In the application of RXCHF to positronic systems, only a single electronic orbital is explicitly correlated to the positronic orbital. We apply NEO-RXCHF to three systems: positron-lithium, lithium positride, and positron-lithium hydride. For all three of these systems, the RXCHF approach provides accurate two-photon annihilation rates, average contact densities, electronic and positronic single-particle densities, and electron-positron contact densities. Moreover, the RXCHF approach is significantly more accurate than the original XCHF approach, in which all electronic orbitals are explicitly correlated to the positronic orbital in the same manner, because the RXCHF wavefunction is optimized to produce a highly accurate description of the short-ranged electron-positron interaction that dictates the annihilation rates and other local properties. Furthermore, RXCHF methods that neglect or approximate the electronic exchange interactions between the geminal-coupled electronic orbital and the regular electronic orbitals lead to virtually identical annihilation rates and densities as the fully antisymmetric RXCHF method but offer substantial advantages in computational tractability. Thus, NEO-RXCHF is a promising, computationally practical approach for studying larger positron-containing systems.


In vitro fertilization and embryo culture in the grey short-tailed opossum, Monodelphis domestica.
In vitro fertilization and early embryo culture was undertaken in the South American marsupial, the grey short-tailed opossum, Monodelphis domestica. Adult females were induced into oestrus by a system of pairing with an unfamiliar male and mature oocytes were recovered from the ovary 15-18 h after mating and placed in pre-warmed modified MEM medium at 33 degrees C (basal body temperature) or 37 degrees C. Spermatozoa recovered from the cauda epididymides of adult males were preincubated in medium for 2 h during which time paired spermatozoa separated and initiated hyperactivated motility. Oocytes were transferred to 0.4 ml drops of spermatozoa containing 0.5-1.0 x 10(6) spermatozoa ml-1. Only single spermatozoa bound to the zona pellucida, and fertilization occurred within 1-2 h as indicated by a breach in the zona and confirmed by electron microscopy. At 37 degrees C, 95 of 152 (62.5%) oocytes were fertilized and 64 (67%) developed to two-cell stage or beyond. At 33 degrees C, 5 of 28 (18%) oocytes were fertilized. This is the first report of complete in vitro fertilization in a marsupial.


Genetic complementation of an outer membrane cytochrome omcB mutant of Shewanella putrefaciens MR-1 requires omcB plus downstream DNA.
Anaerobically grown cells of the metal-reducing bacterium Shewanella putrefaciens MR-1 contain multiple outer membrane (OM) cytochromes. A gene replacement mutant (strain OMCB1) lacking the OM cytochrome OmcB is markedly deficient in the reduction of MnO2 and exhibits reduced rates of Fe(III) reduction. The levels of other OM cytochromes are also decreased in OMCB1. Complementation of OMCB1 with wild-type omcB did not restore any of these defects. However, a 21-kb genomic fragment from MR-1, which included omcB and 19 kb of downstream DNA, fully restored MnO2 and Fe(III) reduction and the full complement of OM cytochromes to OMCB1. A 14.7-kb DNA fragment, including omcB and 12 kb of downstream DNA, provided only a modest increase in MnO2 reduction and OM cytochrome content, but it fully restored Fe(III) citrate reduction and partially restored FeOOH reduction. While omcB mRNA was readily detected in this complement, the OmcB protein was not detected in any cellular compartment. The restoration of Fe(III) reduction despite the absence of OmcB suggests that OmcB itself is not required for Fe(III) reduction. Another OM cytochrome, OmcA, was mislocalized to the cytoplasmic membrane of OMCB1. Only the 21-kb genomic fragment was able to restore proper localization of OmcA to the OM. This 21-kb fragment does not contain omcA, but it does contain several open reading frames (ORFs) downstream from omcB. The most downstream of these ORFs (altA) encodes a putative AraC-like transcriptional regulator. However, a gene replacement mutant of altA resembled the wild type with respect to MnO2 reduction, OM cytochrome content, and the localization of OmcA and OmcB to the OM. Since OMCB1 continues to express genes immediately downstream from omcB, the lack of expression of this downstream DNA does not explain its phenotype or the need for the large complementing fragment. The results suggest that the DNA downstream of omcB must be present in cis in order to restore Fe(III) reduction, MnO2 reduction, OM cytochrome content, and the localization of OmcA and OmcB to the OM.


Another reason to restrict the use of a hypertonic, glucose-based peritoneal dialysis fluid: its impact on peritoneal macrophage function in vivo.
Previous in vitro studies have revealed that the currently used peritoneal dialysis fluids (PDFs) inhibit several functions of phagocytic cells. Glucose-mediated hypertonicity was demonstrated to be one of the major detrimental factors. To investigate the clinical relevance of these in vitro findings, we compared the in vivo effect of a PDF with 3.86% glucose (D386) and a PDF containing 1.36% of glucose (D136) on peritoneal macrophage (PMO) function in eight CCPD patients. In a randomized cross-over setting, each patient used exclusively D136 or D386 on day one. The next day the patients who used D136 were switched to D386 and vice versa. PMO were isolated from the effluents and studied for their phagocytic capacity and chemiluminescence response. PMOs obtained from D386 effluents were significantly less phagocytic as compared to PMOs obtained after the in vivo use of D136 (38 +/- 7.1 vs 63 +/- 8.3%, p less than 0.02). In addition PMOs exposed to D386 were less able to mount a respiratory burst (peak response 4 +/- 1.0 vs 15 +/- 1.4% of control cells, p less than 0.05). Thus, the in vivo use of D386 was accompanied by significantly decreased PMO functions. These findings further stress the need to search for alternative solutions that do not rely on glucose-mediated hypertonicity for ultrafiltration.


Ghrelin receptors in non-Mammalian vertebrates.
The growth hormone secretagogue-receptor (GHS-R) was discovered in humans and pigs in 1996. The endogenous ligand, ghrelin, was discovered 3 years later, in 1999, and our understanding of the physiological significance of the ghrelin system in vertebrates has grown steadily since then. Although the ghrelin system in non-mammalian vertebrates is a subject of great interest, protein sequence data for the receptor in non-mammalian vertebrates has been limited until recently, and related biological information has not been well organized. In this review, we summarize current information related to the ghrelin receptor in non-mammalian vertebrates.


Hypogammaglobulinemia in infants infected with human immunodeficiency virus.
Two premature infants were infected with HIV via blood transfusions during the neonatal period. Although neither patient had serum antibody to HIV owing to severe hypogammaglobulinemia, HIV infection was confirmed by finding HIV antigen in the sera of both patients. These cases show that HIV infection can produce severe hypogammaglobulinemia, and illustrate the value of HIV antigen detection in the diagnosis of HIV infection in seronegative patients.


Combined electrocardiogram and photoplethysmogram measurements as an indicator of objective sleepiness.
There is considerable interest in unobtrusive and portable methods of monitoring sleepiness outside the laboratory setting. This study evaluates the usefulness of combined electrocardiogram (ECG) and photoplethysmogram (PPG) measurements for estimating psychomotor vigilance. The psychomotor vigilance test (PVT) was performed at various points over the course of a day, and one channel each of ECG and PPG was recorded simultaneously. Features derived from ECG and PPG were entered into multiple linear regression models to estimate PVT values. A double-loop, subject-independent validation scheme was used to develop and validate the models. We show that features obtained from the RR interval were reasonably useful for estimating absolute PVT levels, but were somewhat inadequate for estimating within-subject PVT changes. Combined ECG and PPG measurements appear to be useful for predicting PVT values, and deserve further investigation for portable sleepiness monitoring.


Virtual reality-based attention training in patients with neurological damage: A pilot study.
Virtual Reality has been shown to be a valid tool to assess cognitive functions in an ecologically valid way. However, evidence regarding its effectiveness as a treatment option for cognitive rehabilitation has been limited. Furthermore, its potential to facilitate the transfer of training effects to patients' everyday life is still poorly studied. This study aimed to evaluate the efficacy of a VR-based attention training and its transfer to attentional functioning in everyday life. Nineteen inpatients with neurological disorders and attentional deficits underwent daily 30-min VR training sessions over a two-week period. Attentional functions were assessed before and after the training period using several computerized tests, two self-constructed behavioral tasks, and a questionnaire assessing patients' subjective attentional functioning. Pre-Post-analyses show significant decreases in reaction times in the computerized alertness and selective attention tests. Transfer to the behavioral tasks and self-report data could not be observed. Despite the specificity of the changes, confounding effects cannot entirely be ruled out due to the lack of a control group. Results suggest that training was effective in improving attentional functioning along neuropsychological measures, but did not elicit transfer to an ecologically valid or subjective level. Implications for the future development of VR interventions are discussed.


An in vitro model for the regulation of human cytomegalovirus latency and reactivation in dendritic cells by chromatin remodelling.
Human cytomegalovirus (HCMV) is a frequent cause of major disease following primary infection or reactivation from latency in immunocompromised patients. Infection of non-permissive mononuclear cells is used for analyses of HCMV latency in vitro. Using this approach, it is shown here that repression of lytic gene expression following experimental infection of CD34+ cells, a site of HCMV latency in vivo, correlates with recruitment of repressive chromatin around the major immediate-early promoter (MIEP). Furthermore, long-term culture of CD34+ cells results in carriage of viral genomes in which the MIEP remains associated with transcriptionally repressive chromatin. Finally, specific differentiation of long-term cultures of infected CD34+ cells to mature dendritic cells results in acetylation of histones bound to the MIEP, concomitant loss of heterochromatin protein 1 and the reactivation of HCMV. These data are consistent with ex vivo analyses of latency and may provide a model for further analyses of the mechanisms involved during latency and reactivation.


The canonical Wnt signaling pathway plays an important role in lymphopoiesis and hematopoiesis.
The evolutionarily conserved canonical Wnt-beta-catenin-T cell factor (TCF)/lymphocyte enhancer binding factor (LEF) signaling pathway regulates key checkpoints in the development of various tissues. Therefore, it is not surprising that a large body of gain-of-function and loss-of-function studies implicate Wnt-beta-catenin signaling in lymphopoiesis and hematopoiesis. In contrast, recent papers have reported that Mx-Cre-mediated conditional deletion of beta-catenin and/or its homolog gamma-catenin (plakoglobin) did not impair hematopoiesis or lymphopoiesis. However, these studies also report that TCF reporter activity remains active in beta-catenin- and gamma-catenin-deficient hematopoietic stem cells and all cells derived from these precursors, indicating that the canonical Wnt signaling pathway was not abrogated. Therefore, these studies in fact show that the canonical Wnt signaling pathway is important in hematopoiesis and lymphopoiesis, even though the molecular basis for the induction of the reporter activity is currently unknown. In this perspective, we provide a broad background to the field with a discussion of the available data and create a framework within which the available and future studies may be evaluated.


Remote magnetic targeting of iron oxide nanoparticles for cardiovascular diagnosis and therapeutic drug delivery: where are we now?
Magnetic resonance imaging (MRI) allows for an accurate assessment of both functional and structural cardiac parameters, and thereby appropriate diagnosis and validation of cardiovascular diseases. The diagnostic yield of cardiovascular MRI examinations is often increased by the use of contrast agents that are almost exclusively based on gadolinium compounds. Another clinically approved contrast medium is composed of superparamagnetic iron oxide nanoparticles (IONs). These particles may expand the field of contrast-enhanced cardiovascular MRI as recently shown in clinical studies focusing on acute myocardial infarction (AMI) and atherosclerosis. Furthermore, IONs open up new research opportunities such as remote magnetic drug targeting (MDT). The approach of MDT relies on the coupling of bioactive molecules and magnetic nanoparticles to form an injectable complex. This complex, in turn, can be attracted to and retained at a desired target inside the body with the help of applied magnetic fields. In comparison to common systemic drug applications, MDT techniques promise both higher concentrations at the target site and lower concentrations elsewhere in the body. Moreover, concurrent or subsequent MRI can be used for noninvasive monitoring of drug distribution and successful delivery to the desired organ in vivo. This review does not only illustrate the basic conceptual and biophysical principles of IONs, but also focuses on new research activities and achievements in the cardiovascular field, mainly in the management of AMI. Based on the presentation of successful MDT applications in preclinical models of AMI, novel approaches and the translational potential of MDT are discussed.


Baylisascariasis.
The raccoon roundworm, Baylisascaris procyonis, is the most common and widespread cause of clinical larva migrans in animals. In addition, it is increasingly recognized as a cause of devastating or fatal neural larva migrans in infants and young children and ocular larva migrans in adults. Humans become infected by accidentally ingesting infective B. procyonis eggs from raccoon latrines or articles contaminated with their feces. Two features distinguish B. procyonis from other helminthes that cause larva migrans: (i) its aggressive somatic migration and invasion of the central nervous system and (ii) the continued growth of larvae to a large size within the central nervous system. Typically, B. procyonis neural larva migrans presents as acute fulminant eosinophilic meningoencephalitis. Once invasion of the central nervous system has occurred, the prognosis is grave with or without treatment. To date, despite anthelmintic treatment of cases of B. procyonis neural larva migrans, there are no documented neurologically intact survivors. Epidemiologic study of human cases of neural larva migrans demonstrate that contact with raccoon feces or an environment contaminated by infective eggs and geophagia or pica are the most important risk factors for infection. In many regions of the United States, increasingly large populations of raccoons, with high rates of B. procyonis infection, live in close proximity to humans. Although documented cases of human baylisascariasis remain relatively uncommon, widespread contamination of the domestic environment by infected raccoons suggests that the risk of exposure and human infection is probably substantial. In the absence of early diagnosis or effective treatment, prevention of infection is the most important public health measure.


Evaluation of oxygen partial pressure, temperature and stripping of antioxidants for accelerated shelf-life testing of oil blends using 1H NMR.
Lipid oxidation compromises the shelf-life of lipid-containing foods, leading to the generation of unpleasant off-flavours. Monitoring lipid oxidation under normal shelf-life conditions can be time-consuming (i.e. weeks or months) and therefore accelerated shelf-life conditions are often applied. However, little is known on their impact on the lipid oxidation mechanisms. In this study, different oxygen partial pressures (PO2; 10 and 21%), temperatures (20, 30 and 40 °C), and the removal of antioxidants through stripping of the oil were tested to accelerate lipid oxidation. Increasing the incubation temperature of stripped oil blends from 30 to 40 °C reduced the onset of lipid oxidation from 4 to 2 weeks, whereas the PO2 had no impact. Surprisingly, at room temperature, an increase in PO2 resulted in a longer onset time (10 weeks under 10% oxygen, 15 weeks under 21% oxygen). We hypothesize that this is due to a shift in (initiation) mechanism. In non-stripped oil, an increase in PO2 from 10 to 21% decreased the onset time from 16 to 10 weeks (40 °C). Temperature elevations and stripping led to a shift towards more trans-trans diene hydroperoxides, as compared to the cis-trans conformation. Additionally, oil stripping led to an increase in oxidized PUFAs with three or more double bonds in which the hydroperoxide group is located between the double bond pattern, instead of on the edge of it. Lastly, it was shown that small additions of LC-PUFAs (0, 0.3, 0.6, 1.2 and 2.3%, w/w) accelerate lipid oxidation, even in relatively stable stripped oils. In conclusion, increased PO2 and slightly elevated temperatures hold fair potential for accelerated shelf-life testing of non-stripped oils with a limited impact on the lipid oxidation mechanisms, whereas stripping significantly changes propagation mechanisms.


Accumulation of advanced glycation end products and chronic complications in ESRD treated by dialysis.
Cardiovascular and connective tissue disorders are very common in patients with end-stage renal disease (ESRD), and the accumulation of advanced glycation end products (AGEs) is significantly increased in these patients. Accumulation of AGEs is believed to have a role in tissue protein aging and the pathogenesis of such age-related diseases as diabetes and ESRD. AGEs accumulate in patients with ESRD as a result of nonenzymatic glycation, oxidative stress, and diminished clearance of AGE precursors. Some AGEs show characteristic brown pigmentation and fluorescence, form protein-protein cross-links, and may ligate with AGE-specific receptors, inducing oxidative stress and cytokine production. This review focuses on the clinical relevance of AGE accumulation in patients with ESRD treated by dialysis for the development of long-term complications. The formation and accumulation of AGEs in patients with ESRD are discussed, as well as the relationship between AGE accumulation and such major complications of ESRD as cardiovascular and connective tissue disorders.


Ribosome-dependent activation of stringent control.
In order to survive, bacteria continually sense, and respond to, environmental fluctuations. Stringent control represents a key bacterial stress response to nutrient starvation that leads to rapid and comprehensive reprogramming of metabolic and transcriptional patterns. In general, transcription of genes for growth and proliferation is downregulated, while those important for survival and virulence are upregulated. Amino acid starvation is sensed by depletion of the aminoacylated tRNA pools, and this results in accumulation of ribosomes stalled with non-aminoacylated (uncharged) tRNA in the ribosomal A site. RelA is recruited to stalled ribosomes and activated to synthesize a hyperphosphorylated guanosine analogue, (p)ppGpp, which acts as a pleiotropic secondary messenger. However, structural information about how RelA recognizes stalled ribosomes and discriminates against aminoacylated tRNAs is missing. Here we present the cryo-electron microscopy structure of RelA bound to the bacterial ribosome stalled with uncharged tRNA. The structure reveals that RelA utilizes a distinct binding site compared to the translational factors, with a multi-domain architecture that wraps around a highly distorted A-site tRNA. The TGS (ThrRS, GTPase and SpoT) domain of RelA binds the CCA tail to orient the free 3' hydroxyl group of the terminal adenosine towards a β-strand, such that an aminoacylated tRNA at this position would be sterically precluded. The structure supports a model in which association of RelA with the ribosome suppresses auto-inhibition to activate synthesis of (p)ppGpp and initiate the stringent response. Since stringent control is responsible for the survival of pathogenic bacteria under stress conditions, and contributes to chronic infections and antibiotic tolerance, RelA represents a good target for the development of novel antibacterial therapeutics.


Psychosocial mediators of antiretroviral nonadherence in HIV-positive adults with substance use and mental health problems.
This study used data from 1,889 HIV-positive patients on antiretroviral (ARV) medications who participated in the HIV Cost and Services Utilization Study to investigate whether nonadherence to ARV medications among patients with mental health and substance use problems could be explained by difficulty getting and negative attitudes toward ARV medications, poor fit of the regimen with lifestyle, lack of instruction and cues for remembering the regimen from a health care provider, and poor support from others for taking ARV medications. Difficulty getting ARV medications and poor fit with lifestyle were significant mediators of nonadherence for patients with a probable psychiatric disorder. Difficulty getting medication was a mediator for heavy drinkers, and poor fit with lifestyle was a mediator for drug users who drank heavily. Further research is needed to identify and address the barriers to adherence in these populations.


Benzene metabolism by human liver microsomes in relation to cytochrome P450 2E1 activity.
Low levels of benzene from sources including cigarette smoke and automobile emissions are ubiquitous in the environment. Since the toxicity of benzene probably results from oxidative metabolites, an understanding of the profile of biotransformation of low levels of benzene is critical in making a valid risk assessment. To that end, we have investigated metabolism of a low concentration of [14C]benzene (3.4 microM) by microsomes from human, mouse and rat liver. The extent of phase I benzene metabolism by microsomal preparations from 10 human liver samples and single microsomal preparations from both mice and rats was then related to measured activities of cytochrome P450 (CYP) 2E1. Measured CYP 2E1 activities, as determined by hydroxylation of p-nitrophenol, varied 13-fold (0.253-3.266 nmol/min/mg) for human samples. The fraction of benzene metabolized in 16 min ranged from 10% to 59%. Also at 16 min, significant amounts of oxidative metabolites were formed. Phenol was the main metabolite formed by all but two human microsomal preparations. In those samples, both of which had high CYP 2E1 activity, hydroquinone was the major metabolite formed. Both hydroquinone and catechol formation showed a direct correlation with CYP 2E1 activity over the range of activities present. A simulation model was developed based on a mechanism of competitive inhibition between benzene and its oxidized metabolites, and was fit to time-course data for three human liver preparations. Model calculations for initial rates of benzene metabolism ranging from 0.344 to 4.442 nmol/mg/min are directly proportional to measured CYP 2E1 activities. The model predicted the dependence of benzene metabolism on the measured CYP 2E1 activity in human liver samples, as well as in mouse and rat liver samples. These results suggest that differences in measured hepatic CYP 2E1 activity may be a major factor contributing to both interindividual and interspecies variations in hepatic metabolism of benzene. Validation of this system in vivo should lead to more accurate assessment of the risk of benzene's toxicity following low-level exposure.


Inhibition of Neutrophil Secretion Upon Adhesion as a Basis for the Anti-Inflammatory Effect of the Tricyclic Antidepressant Imipramine.
Recent studies demonstrate the involvement of inflammatory processes in the development of depression and the anti-inflammatory effects of antidepressants. Infiltration and adhesion of neutrophils to nerve tissues and their aggressive secretion are considered as possible causes of inflammatory processes in depression. We studied the effect of the antidepressant imipramine on the adhesion and accompanied secretion of neutrophils under control conditions and in the presence of lipopolysaccharides (LPS). As a model of integrin-dependent neutrophil infiltration into tissues, we used integrin-dependent adhesion of neutrophils to the fibronectin-coated substrate. Imipramine inhibited neutrophil adhesion and concomitant secretion of proteins, including matrix metalloproteinase 9 (MMP-9) and neutrophil gelatinase-associated lipocalin (NGAL), which modify the extracellular matrix and basement membranes required for cell migration. Imipramine also significantly and selectively blocked the release of the free amino acid hydroxylysine, a product of lysyl hydroxylase, an enzyme that affects the organization of the extracellular matrix by modifying collagen lysine residues. In contrast, imipramine enhanced the release of ROS by neutrophils during adhesion to fibronectin and stimulated apoptosis. The anti-inflammatory effect of imipramine may be associated with the suppression of neutrophil infiltration and their adhesion to nerve tissues by inhibiting the secretion of neutrophils, which provides these processes.


Crosstalk between cGAS-STING pathway and autophagy in cancer immunity.
The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway is critical in cancer immunity. Autophagy is a highly conserved process that is responsible for the degradation of cytoplasmic material and is involved in both innate and adaptive immunity. Recently, cGAS-STING and autophagy have been shown to be interconnected, which may influence the progression of cancer. Although cGAS-STING and autophagy have been shown to be interrelated in innate immunity, little has been reported about cancer immunity. As cancer immunity is key to treating tumors, it is essential to summarize the relationship and interactions between the two. Based on this, we systematically sorted out the recent findings of cGAS-STING and autophagy in cancer immunity and explored the interactions between cGAS-STING and autophagy, although these interactions have not been extensively studied. Lastly, we provide an outlook on how cGAS-STING and autophagy can be combined, with the hope that our research can help people better understand their potential roles in cancer immunity and bring light to the treatment of cancer.


Phase I study of the 177Lu-DOTA0-Tyr3-Octreotate (lutathera) in combination with nivolumab in patients with neuroendocrine tumors of the lung.
Lutathera is a 177Lutetium-labeled somatostatin analog approved for the treatment of gastroenteropancreatic neuroendocrine tumors (NETs). Somatostatin receptors are expressed in small cell lung cancer (SCLC). Nivolumab, an anti-PD-1 antibody, may act synergistically with lutathera to generate antitumor immunity. We conducted a phase I study of lutathera plus nivolumab in patients with advanced NETs of the lung. Patients with relapsed/refractory extensive-stage SCLC (ES-SCLC), non-progressing ES-SCLC after first-line platinum-based chemotherapy, or advanced grade I-II pulmonary NETs were eligible. The primary objective was to determine the recommended phase 2 dose (RP2D). The phase I portion followed a standard 3+3 design, assessing two dose levels (dose level 1: lutathera 3.7 GBq every 8 weeks for four doses with nivolumab 240 mg every 2 weeks; dose level 2: lutathera 7.4 GBq every 8 weeks for four doses with nivolumab 240 mg every 2 weeks). Nine patients were enrolled (six ES-SCLC, two pulmonary atypical carcinoid, one high-grade pulmonary neuroendocrine carcinoma). No dose-limiting toxicities (DLTs) were observed at dose level 1. At dose level 2, one patient with refractory ES-SCLC developed a DLT (grade 3 rash). The most common treatment-related adverse events (TRAEs) were lymphopenia (n=7), thrombocytopenia (n=4), anemia (n=3), and nausea (n=3). The most common grade 3 TRAE was lymphopenia (n=4). Among the seven patients with measurable disease, one patient with ES-SCLC had a partial response. Two patients with pulmonary atypical carcinoid had stable disease lasting 6 months. The RP2D was dose level 2. Lutathera plus nivolumab was well tolerated and showed signs of antitumor activity. This combination warrants further exploration. NCT03325816.


Biosynthesis of human colonic mucin: Muc2 is the prominent secretory mucin.
Human colonic epithelium produces large amounts of mucin. The aim of this study was to examine mucin biosynthesis in the human colon. Human colonic mucin was isolated using CsCl density gradients, and polyclonal antiserum was raised. Biosynthesis of colonic mucins was studied by labeling colonic explants with 35S-labeled amino acids or [35S]sulfate and subsequent immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The polyclonal antiserum specifically recognized colonic mucin, primarily reacting with peptide epitopes. Biosynthetic pulse/chase experiments showed a 35S-amino acid-labeled mucin precursor of about 600 kilodaltons, which was converted into a mature, glycosylated, and sulfated mucin and subsequently secreted into the medium. This mature mucin comigrated with isolated colonic mucin with an apparent molecular weight of 550 kilodaltons on SDS-PAGE, whereas gel filtration indicated that the molecular weight is actually much larger. Independent immunoprecipitation with an anti-Muc2 antiserum showed cross-reactivity with the 600-kilodalton precursor. These results show the biosynthesis of a secretory colonic mucin for the first time. This mucin is synthesized as a precursor protein of approximately 600 kilodaltons, which, after glycosylation, is secreted as a glycoprotein with an apparent molecular weight of 550 kilodaltons on SDS-PAGE. It is very likely that this mucin is Muc2.